Enzyme responsive nanocapsules for protein delivery

Patent metadata

Field	Value
Publication number	US-9993440-B2
Application number	US-201214342524-A
Country	US
Kind code	B2
Filing date	Sep 4, 2012
Priority date	Sep 2, 2011
Publication date	Jun 12, 2018
Grant date	Jun 12, 2018

Abstract

The invention provides methods of making and using compositions comprising a polymer shell designed to deliver polypeptides to selected environments. In embodiments of the invention, different environmental conditions are harnessed to allow the selective degradation of the polymer shell and the consequential release of one or polypeptides encapsulated therein. In illustrative embodiments, polymer components of the shell are interconnected by peptide-containing crosslinker moieties, linkages which maintain the integrity of the polymer shell under certain environmental conditions, but can also be cleaved when combined with a selected protease.

First claim

The invention claimed is:

1. A composition of matter comprising:
at least one polypeptide;
a polymeric network, wherein:
polymers in the polymeric network are coupled together by a first crosslinking agent and a second crosslinking agent so as to form a shell that encapsulates the polypeptide;
the first crosslinking agent comprises a peptide having an amino acid sequence that is cleaved by a protease, wherein amino acid sequence that is cleaved by the protease comprises KNRVK (SEQ ID NO: 1), GGIPVSLRSGGK (SEQ ID NO: 2) or GGVPLSLYSGGK (SEQ ID NO: 3); and
the second crosslinking agent does not comprise a peptide having an amino acid sequence that is cleaved by a protease;
wherein the polymers, the first crosslinking agent and the second crosslinking agent are disposed within the polymeric network in an orientation so that proteolytic cleavage of the first crosslinking agent releases the polypeptide from the shell into an external environment.

2. The composition of claim 1 , wherein relative molar amounts of the first crosslinking agent and the second crosslinking agent in the shell are at least 1:5, 1:4, 1:3, 1:2, 1:1, 2:1, 3:1, 4:1 or 5:1.

3. The composition of claim 1 , wherein shell that encapsulates the polypeptide has a diameter between 15 and 35 nanometers.

4. The composition of claim 1 , wherein the polypeptide comprises a protein that stimulates cellular growth.

5. The composition of claim 1 , wherein the composition further comprises a hydrogel in which the shell is disposed.

6. The composition of claim 5 , wherein the shell is covalently coupled to the hydrogel by a hydrogel crosslinking agent.

7. The composition of claim 6 , wherein the hydrogel crosslinking agent comprises a peptide having an amino acid sequence that is cleaved by a protease.

8. The composition of claim 1 , wherein the first crosslinking agent comprises a peptide having D-amino acids.

9. The composition of claim 1 , wherein the concentration of the first crosslinking agent in the composition relative to the concentration of the second crosslinking agent in the composition is controlled so as to modulate the rate at which the polypeptide is released from the shell into an external in vivo environment.

10. The composition of claim 9 , wherein the concentration of the first crosslinking agent is greater than the concentration of the second crosslinking agent in order to modulate the rate at which the polypeptide is released from the shell into an external in vivo environment.

11. The composition of claim 9 , wherein the concentration of the first crosslinking agent is less than the concentration of the second crosslinking agent in order to modulate the rate at which the polypeptide is released from the shell into an external in vivo environment.

12. A composition of matter comprising:
at least two polypeptides;
a polymeric network, wherein:
polymers in the polymeric network are coupled together by a first crosslinking agent and a second crosslinking agent so as to form a shell that encapsulates the at least two polypeptides;
the first crosslinking agent comprises a peptide having an amino acid sequence that is cleaved by a protease, wherein amino acid sequence that is cleaved by the protease comprises KNRVK (SEQ ID NO: 1), GGIPVSLRSGGK (SEQ ID NO: 2) or GGVPLSLYSGGK (SEQ ID NO: 3); and
the second crosslinking agent does not comprise a peptide having an amino acid sequence that is cleaved by the protease;
wherein the polymers, the first crosslinking agent and the second crosslinking agent are disposed within the polymeric network in an orientation so that proteolytic cleavage of the first crosslinking agent releases the at least two polypeptides from the shell into an external environment.

Assignees

• Lu Yunfeng
• Segura Tatiana
• Zhu Suwei
• Wen Jing
• Univ California

Inventors

• Lu Yunfeng
• Segura Tatiana
• Zhu Suwei
• Wen Jing

Classifications

• A61K9/5192

  Processes · CPC title

• A61K47/65

  Peptidic linkers, binders or spacers, e.g. peptidic enzyme-labile linkers · CPC title

• A61K9/19

  lyophilised {, i.e. freeze-dried, solutions or dispersions (lyophilised products with subsequent particle size reduction A61K9/14; granules or pellets made by lyphilisation A61K9/1682; solid oral dosage forms made by lyophilisation A61K9/2095; lyophilisation additives A61K47/00)} · CPC title

• A61K9/5169 Primary

  Proteins, e.g. albumin, gelatin · CPC title

• A61K9/5138

  obtained by reactions only involving carbon-to-carbon unsaturated bonds, e.g. polyvinyl pyrrolidone, poly(meth)acrylates · CPC title